JP6476253B2 - N- (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl-4- ( 2-Oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate salt - Google Patents

Description

(Cross-reference of related applications)
This application claims the benefit of US Provisional Application No. 61 / 603,598, filed February 27, 2012.

  N- (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl-4- ( Disclosed is the hemisulfate salt of 2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate and its crystalline form. N- (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl-4- ( At least one pharmaceutical composition comprising 2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate hemisulfate and a CGRP related disorder (e.g. N- (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] At least one hemisulfate of pyridin-9-yl-4- (2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate Use methods are also disclosed.

（Ｉ）
の構造を有し、本明細書では「化合物（Ｉ）」と称される。化合物（Ｉ）、化合物（Ｉ）の製造方法、および化合物（Ｉ）を用いる治療方法は、特許文献１に開示されている。この文献は、本出願人に譲渡されており、その全体が出典明示により本明細書に取り込まれるものである。 Compound, N- (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl-4 -(2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate has the formula I:

(I)
And is referred to herein as “Compound (I)”. Patent Document 1 discloses Compound (I), a method for producing Compound (I), and a therapeutic method using Compound (I). This document is assigned to the applicant and is incorporated herein in its entirety by reference.

  The usefulness of oral formulations depends, inter alia, on the degree of consistency in bioavailability where the active agent has bioavailability between patients. The bioavailability of orally administered drugs is often affected by a variety of factors including, for example, drug solubility in the gastrointestinal tract, drug stability in the gastrointestinal tract, and drug absorption in the gastrointestinal tract. Furthermore, these factors can be affected by co-administration of other drugs and / or food intake, resulting in variations in the bioavailability of orally administered drugs. In addition, rapid in vivo dissolution of the active agent is also required to provide a rapid treatment of symptoms such as migraine.

The dissolution rate of compound (I) depends on the pH of the aqueous medium. Compound (I) has a higher dissolution rate at a pH value of 1-5 than a pH value of 7. In oral administration of Compound (I), the dissolution rate and Compound I bioavailability can be affected by the pH of the gastric contents. The normal pH of the stomach is 1.2 to 1.8 according to Non-Patent Document 1. However, patients, antacids which can reduce the dissolution rate of the compound (I), proton pump inhibitors, and H _2, such as famotidine - other containing receptor antagonists, can raise the pH of the stomach You may also take some medications.

  Typically, in the manufacture of pharmaceutical compositions, the active ingredient forms are sought that have the desired balance of properties, such as, for example, dissolution rate, solubility, bioavailability, and / or storage stability. For example, a form that is sufficiently soluble and bioavailable has another form that has an undesirable solubility and / or bioavailability profile during manufacturing, preparation, and / or storage of the pharmaceutical composition. Active ingredient forms are sought that have sufficient stability, solubility, and bioavailability to prevent conversion. For example, search for active ingredient forms that are stable at ambient temperature and humidity conditions and have low hygroscopicity.

  Furthermore, forms of active ingredients that can be produced by methods capable of producing active ingredients on a large scale can also be explored. In such methods, it is desirable that the active ingredient be in a form that can be easily isolated and / or purified (eg, by filtration) and dried.

  Furthermore, since production economics are important, it is desirable to avoid using costly materials in the production of forms.

  Applicants have surprisingly reduced the bioavailability of Compound (I), matched bioavailability between patients, and / or increased the bioavailability of Compound (I) to patients. Found hemisulfate. Applicants also surprisingly have reduced compounds (I) bioavailability variability, matched bioavailability between patients and / or increased compound (I) bioavailability to patients (I ) Was found. The hemisulfate salt of compound (I) and its crystalline form surprisingly has a balance of properties required in a pharmaceutical composition. The invention also relates to other important aspects.

US Patent Publication No. 2011 / 0251223A1

C.J.Perigard, Clinical Analysis, Chapter 32, in Remington: The Science and Practice of Pharmacy 20th Edition, A.R.Gennaro, editor; 2000, Lippinocott Williams & Wilkins, Baltimore, MD

（Ｉ）
のヘミ硫酸塩である。 In certain embodiments, the present invention provides compound (I):

(I)
Of hemisulfate.

  The present invention also provides a crystalline form of the hemisulfate salt of Compound (I).

  The present invention also provides a pharmaceutical composition comprising a pharmaceutically acceptable carrier and / or diluent; and a hemisulfate salt of Compound (I).

  The present invention also provides a method for treating a disease or disorder associated with the activity of the CGRP receptor, which comprises administering a hemisulfate salt of Compound (I) to a mammalian patient.

  The present invention also provides methods and intermediates for preparing the hemisulfate salt of Compound (I) and / or its crystalline form.

  The present invention also provides a hemisulfate salt of Compound (I) for use in therapy.

  The present invention also provides a therapeutic agent for migraine, neurogenic vasodilation, neurogenic inflammation, burns, circulatory shock, flushing associated with menopause, airway inflammatory diseases such as asthma, and chronic obstructive pulmonary disease (COPD). Provided is the hemisulfate salt of Compound (I) for manufacture.

  These and other features of the invention are described in an expanded manner as the disclosure continues.

FIG. 1 shows an experimental (room temperature) and simulation (room temperature) PXRD pattern (CuKα λ = 1.5418 Å) of Form H1.5-1 of Example Compound 1. FIG. 2 shows the differential scanning calorimetry profile of Example Compound 1 Form H1.5-1. FIG. 3 shows the thermogravimetric analysis profile of Example Compound 1 Form H1.5-1. FIG. 4 shows the cross-polarization magic angle rotation (CPMAS) NMR spectrum of Example Compound 1 Form H1.5-1. FIG. 5 shows the moisture absorption isotherm for Example Compound 1 at 25 ° C. (X) Adsorption; (■) Desorption. FIG. 6 shows a pH value of about 5 in simulated intestinal simulated intestinal fluid (FeSSIF) and fasted intestinal simulated intestinal fluid (FaSSIF) for Compound (I) and the HCl salt of Compound (I) as the free base. The% solubility over time at a pH value of about 7 is shown. FIG. 7 shows a pH value of about 1 for compound (I) and hemisulfate of compound (I) as the free base, a pH value of about 5 in satiety small intestinal simulated intestinal fluid (FeSSIF), and fasting Figure 6 shows% solubility over time at a pH value of about 7 in simulated small intestinal fluid (FaSSIF). FIG. 8 shows the plasma pharmacokinetics of compound (I), the free base in humans after oral administration with or without pretreatment with famotidine (40 mg) 2 hours before administration of compound (I). Compound (I) was administered at a dose of 150 mg. (●) Compound (I) (nM); (♦) Compound (I) (nM) by famotidine pretreatment. The x-axis is time (minutes). FIG. 9 shows plasma pharmacokinetics of Compound (I) in dogs after oral administration of Compound (I) and Example Compound 1. Compound (I) and Example Compound 1 were orally administered at a dose of 150 mg (or equivalent). (◆) Pretreatment with compound (I) (nM) and pentagastrin (6 μm / km); (■) Compound (I) and famotidine (40 mg); (▲) Example compound 1 (hemisulfate) and famotidine ( 40 mg). FIG. 10 shows an experimental (room temperature) PXRD pattern (CuKα λ = 1.5418Å) of Example Compound 1 Form P22C. FIG. 11 shows an experimental (room temperature) PXRD pattern (CuKα λ = 1.5418 Å) of Form P33 of Example Compound 1. FIG. 12 shows an experimental (room temperature) PXRD pattern (CuKα λ = 1.5418 Å) of Form P35 of Example Compound 1.

  The features and advantages of the present invention may be readily understood by those of ordinary skill in the art upon reading the following detailed description. It should be appreciated that certain features of the invention described above and below as separate embodiments for clarity may also be combined to form one embodiment. On the contrary, the various features of the invention described in one embodiment for the sake of brevity can also be combined to form partial combinations thereof.

  The names used herein to characterize a particular form, for example, “H1.5-1”, “P22C”, “P33” and “P35” are interpreted by the characterization information presented herein. It is merely an identifier that should be made and should not be limited to excluding other substances with similar or identical physical and chemical characteristics.

  The definitions set forth herein take precedence over definitions set forth in any patent, patent application, and / or patent application publication incorporated herein by reference.

  All numbers representing the amount of ingredients, weight percent, temperature, etc., with the word “about” preceding, result substantially identical to the numerical value with slight variations above and below the stated numerical value. Should only be understood as approximate values that can be used to achieve Thus, unless indicated to the contrary, numerical parameters preceded by the term “about” are approximations that may vary depending on the desired characteristics that are desired to be obtained. Rather than trying to limit the application of the doctrine to the scope of the claims at least, each numerical parameter should be interpreted by applying the usual known techniques, at least taking into account the number of significant digits described It is.

  All measurements are subject to experimental error and are within the spirit of the invention.

  The hemisulfate of compound (I) is preferably isolated and purified after its preparation to give (“substantially pure”) hemisulfate of compound (I) or more than 99% or more. A composition containing a weight of preferably 99.5% or more, more preferably 99.9% or more is obtained, and subsequently used or formulated as described herein. Such hemisulfate salts of “substantially pure” Compound (I) are also included herein as part of the present invention.

  As used herein, “polymorph” means a crystalline form that has the same chemical structure but different spatial arrangement of molecules and / or ions that form the crystal.

  As used herein, “amorphous” means a solid form of molecules and / or ions that are not crystalline. Amorphous solids do not show a clear X-ray diffraction pattern with a sharp maximum.

  As used herein, the term “substantially pure crystalline form” refers to a compound that is said to contain at least about 90% by weight of the form (based on the weight of hemisulfate of compound (I)). It means the crystalline form of hemisulfate of I). The term “at least about 90% by weight” does not limit the application of the doctrine to the scope of the claims, but is not limited to the following, for example, based on the weight of the hemisulfate salt of Compound (I): 90, about 91, 91, about 92, 92, about 93, 93, about 94, 94, about 95, 95, about 96, 96, about 97, 97, about 98, 98, about 99, 99, and about 100 % By weight. In addition to the hemisulfate of compound (I), other forms of hemisulfate of compound (I) (including amorphous hemisulfate of compound (I)) and / or reaction impurities and / or, for example, Processing impurities that occur when the hemisulfate is prepared and / or the crystalline form is prepared may be included.

  The presence of reaction impurities and / or processing impurities can be determined by analytical techniques known in the art, such as chromatography, nuclear magnetic resonance spectroscopy, mass spectrometry, and / or infrared spectroscopy.

  As used herein, the parameter “molecule / asymmetric unit” means the number of molecules of compound (I) in the asymmetric unit.

  As used herein, the unit cell parameter “molecule / unit cell” means the number of molecules of compound (I) in the unit cell.

The present invention is intended to include all isotopes of atoms present in the present compounds. Isotopes include those atoms having the same atomic number but different mass numbers. For general illustration and not limitation, isotopes of hydrogen include deuterium and tritium. Carbon isotopes include ¹³ C and ¹⁴ C. The isotopically-labeled compounds of the present invention are generally prepared using conventional techniques known to those skilled in the art or methods described herein, using appropriately isotope-labeled reagents in place of unlabeled reagents used elsewhere. Can be prepared by methods analogous to. Such compounds can have a variety of potential uses, for example, as standards and reagents in measuring biological activity. In the case of stable isotopes, such compounds may have the potential to preferably modify biological, pharmacological or pharmacokinetic properties.

（Ｉ）
のヘミ硫酸塩を提供する。化合物（Ｉ）のヘミ硫酸塩は、化合物（Ｉ）の各分子に対して０．５のＨ_２ＳＯ_４分子の比率で有する化合物（Ｉ）の酸塩であり、名称：（５Ｓ，６Ｓ，９Ｒ）−５−アミノ−６−（２，３−ジフルオロフェニル）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−９−イル４−（２−オキソ−２，３−ジヒドロ−１Ｈ−イミダゾ［４，５−ｂ］ピリジン−１−イル）ピペリジン−１−カルボキシレート，ヘミ硫酸塩を有する。 The first aspect of the present invention is the compound (I)

(I)
Of hemisulfate. The hemisulfate of compound (I) is an acid salt of compound (I) having a ratio of 0.5 molecule of H ₂ SO ₄ to each molecule of compound (I), and the name: (5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl 4- (2-oxo-2,3- Dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate, hemisulfate.

In certain embodiments, the hemisulfate salt of Compound (I) is a sesquihydrate having a ratio of 1.5 water molecules and 0.5 H ₂ SO ₄ molecules for each molecule of Compound (I). Provided.

  In certain embodiments, the hemisulfate salt of Compound (I) is provided as a crystalline form.

In certain embodiments, the hemisulfate salt of Compound (I) is provided as a crystalline form, and the crystalline form is Form H1.5-1. This crystalline form has a ratio of 1.5 water molecules and 0.5 H ₂ SO ₄ molecules for each molecule of compound (I).

In certain embodiments, Form H1.5-1 is:
Grid size:
a = 10.92Å
b = 33.04Å
c = 7.90cm
α = 90 degrees β = 90 degrees γ = 90 degrees Space group: P2 ₁ 2 ₁ 2
Compound (I) molecule / asymmetric unit: 1
Volume = 2851 ^{３ 3}
Density (calculation) = 1.423 g / cm ³
(Measurement of the crystalline form is at a temperature of about 25 ° C.)
Is characterized by unit cell parameters substantially equivalent to

  In certain embodiments, Form H1.5-1 is characterized by an observed powder X-ray diffraction pattern substantially according to the pattern shown in FIG.

  In certain embodiments, Form H1.5-1 is characterized by a simulated powder X-ray diffraction pattern substantially according to the pattern shown in FIG.

  In certain embodiments, Form H1.5-1 is 5.4 ± 0.1, 8.6 ± 0.1, 9.7 ± 0.1, 12.4 ± 0.1, 14.9 ± 0. .4, selected from 1, 17.6 ± 0.1, 18.1 ± 0.1, 20.5 ± 0.1, 21.4 ± 0.1, and 22.0 ± 0.1 Further, preferably characterized by a powder X-ray diffraction pattern (CuKα λ = 1.5418 Å) containing 5 or more 2θ values (measurement of crystal form is at a temperature of about 25 ° C.).

  In another embodiment, Form H1.5-1 is characterized by a solid state nuclear magnetic resonance spectrum (ssNMR) substantially according to the spectrum shown in FIG.

  In certain embodiments, Form H1.5-1 is 26.6 ± 0.1, 27.1 ± 0.1, 28.3 ± 0.1, 30.7 ± 0.1, 43.1 ± 0. .1, 45.9 ± 0.1, 47.1 ± 0.1, 52.0 ± 0.1, 54.2 ± 0.1, 72.5 ± 0.1, 117.0 ± 0.1 117.7 ± 0.1, 124.2 ± 0.1, 125.2 ± 0.1, 128.3 ± 0.1, 130.3 ± 0.1, 131.4 ± 0.1, 134 .1 ± 0.1, 140.8 ± 0.1, 144.7 ± 0.1, 148.7 ± 0.1, 149.8 ± 0.1, 151.2 ± 0.1, 153.4 6 or more, preferably 7 or more peaks selected from ± 0.1, 155.1 ± 0.1, 155.6 ± 0.1, and 156.7 ± 0.1 ( Solid containing δ (ppm) standardized by TMS Characterized by resonance spectrum.

^＊ジフルオロフェニル環は、０．８１７（５）および０．１８３（５）を占有する２つの配向性（Ｆ１／Ｆ１Ａ、Ｆ２／Ｆ２Ａ）により結晶中で不規則であることが測定される。
表２
表２：２５℃で算出される形態Ｈ１．５−１の分率原子座標。
水素原子および等価原子の原子座標（ｘ１０^４）
等方性変位パラメータ（Å^２ｘ１０^３）

In still further embodiments, Form H1.5-1 is characterized by fractional atomic coordinates substantially as set forth in Table 1.
Table 1: fractional atomic coordinates of form H1.5-1 calculated at 25 ° C.
Atomic coordinates of non-hydrogen atoms and equivalent atoms (x10 ⁴ )
Isotropic displacement parameter (Å ² x10 ³ )

^{* The} difluorophenyl ring is measured to be irregular in the crystal due to two orientations (F1 / F1A, F2 / F2A) occupying 0.817 (5) and 0.183 (5).
Table 2
Table 2: Fraction atomic coordinates of Form H1.5-1 calculated at 25 ° C.
Atomic coordinates of hydrogen atoms and equivalent atoms (x10 ⁴ )
Isotropic displacement parameter (Å ² x10 ³ )

  In an even further embodiment, Form H1.5-1 is characterized by a DSC thermogram substantially as shown in FIG.

  In yet another embodiment, Form H1.5-1 is characterized by a TGA thermogram, and Form H1.5-1 is about 4-5% by weight by heating to a temperature of about 200 ° C. Go through loss.

  In still further embodiments, Form H1.5-1 is substantially identical to the TGA thermogram shown in FIG.

  In still yet another embodiment, Form H1.5-1 is provided in substantially pure crystalline form.

  In yet a further embodiment, the hemisulfate salt of Compound (I) is at least about 90%, preferably at least about 95%, more preferably at least about 99% by weight of Form H1 based on hemisulfate. .5-1.

  In an even further embodiment, substantially pure form H1.5-1 is about 10% or less of the total peak area of the PXRD pattern measured experimentally resulting from peaks that are not present in the simulated PXRD pattern, preferably It has a substantially pure phase uniformity of about 5% or less, more preferably about 2% or less. Most preferably, the substantially pure form H1.5-1 is substantially pure of no more than about 1% of the total peak area of the PXRD pattern measured experimentally resulting from peaks that are not present in the simulated PXRD pattern. Has phase uniformity.

  In another embodiment, the hemisulfate salt of Compound (I) consists essentially of Form H1.5-1. The crystalline form of this embodiment may comprise at least about 90% by weight, preferably at least about 95% by weight, more preferably at least about 99% by weight, based on the weight of the hemisulfate salt of Compound (I).

  In yet another embodiment, the pharmaceutical composition comprises a hemisulfate salt of Compound (I) in Form H1.5-1; and at least one pharmaceutically acceptable carrier and / or diluent.

In certain embodiments, the hemisulfate salt of Compound (I) is provided as a crystalline form, wherein the crystalline form is P22C. This crystalline form is a sesquihydrate with a ratio of 1.5 water molecules and 0.5 H ₂ SO ₄ molecules for each molecule of compound (I).

  In certain embodiments, Form P22C is characterized by an observed powder X-ray diffraction pattern substantially according to the powder X-ray diffraction pattern shown in FIG.

In certain embodiments, the hemisulfate salt of Compound (I) is provided as a monohydrate having a ratio of 1 water molecule and 0.5 H ₂ SO ₄ molecule for each molecule of Compound (I).

In certain embodiments, the hemisulfate salt of Compound (I) is provided as a crystalline form that is Form P33. This crystalline form has a ratio of 1 water molecule and 0.5 H ₂ SO ₄ molecule for each molecule of compound (I).

  In certain embodiments, Form P33 is characterized by an observed powder X-ray diffraction pattern substantially according to the powder X-ray diffraction pattern shown in FIG.

  Compound (I) is suitable as a CGRP receptor antagonist, migraine, neurogenic vasodilation, neurogenic inflammation, burns, circulatory shock, flushing associated with menopause, airway inflammatory diseases such as asthma, and chronic obstruction Useful for the treatment of CGRP-related disorders, including congenital lung disease (COPD).

  Calcitonin gene-related peptide (CGRP) is a naturally occurring 37-amino acid peptide first identified in 1982 (Amara, S. G. et al, Science 1982, 298, 240-244). Two forms of this peptide are expressed (αCGRP and βCGRP) that differ in rat and human by 1 and 3 amino acids, respectively. The peptides are widely distributed both in the periphery (PNS) and the central nervous system (CNS) and are primarily localized in sensory afferents and central nerves and exhibit many biological effects including vasodilation.

  When released from cells, CGRP binds to specific cell surface G protein-coupled receptors and exerts its biological effects preferentially by activation of intracellular adenylate cyclase (Poyner, DR et al, Br J Pharmacol 1992, 105, 441-7; Van Valen, F. et al, Neurosci Lett 1990, 119, 195-8.). Two classes of CGRP receptors (CGRP1 and CGRP2) are presumed to activate CGRP2 receptors based on the antagonist properties of peptide fragment CGRP (8-37) and the ability of linear analogs of CGRP (Juaneda, C. et al. TiPS 2000, 21, 432-438). However, there is no molecular evidence for the CGRP2 receptor (Brain, S. D. et al, TiPS 2002, 23, 51-53). CGRP1 receptor has two components: (i) 7-transmembrane calcitocin receptor-like receptor (CRLR); (ii) 1-transmembrane receptor activity regulatory protein 1 (RAMP1); and (iii) It has an intracellular receptor component protein (RCP) (Evans BN et al., J Biol Chem. 2000, 275, 31438-43). RAMP1 is required for transport of CRLR to the cell membrane and for ligands that bind to the CGRP receptor (McLatchie, L. M. et al, Nature 1998, 393, 333-339). RCP is required for signal transduction (Evans B. N. et al., J Biol Chem. 2000, 275, 31438-43). When small molecule antagonists bind to the CGRP receptor, species-specific differences with typical higher affinity seen for antagonism of human receptors than other species are known (Brain, SD et al , TiPS 2002, 23, 51-53). The amino acid sequence of RAMP1 determines species selectivity, in particular the amino acid residue Trp74 is associated with the human receptor phenotype (Mallee et al. J Biol Chem 2002, 277, 14294-8).

  Inhibitors at the receptor level for CGRP are envisioned to be useful in pathophysiological conditions that result in excessive CGRP receptor activation. These include neurogenic vasodilation, neurogenic inflammation, migraine, cluster headache and other headaches, burns, circulatory shock, menopausal flush, and asthma. Activation of the CGRP receptor has been implicated in the pathogenesis of migraine (Edvinsson L. CNS Drugs 2001; 15 (10): 745-53; Williamson, DJ Microsc. Res. Tech. 2001, 53, 167- 178 .; Grant, AD Brit. J. Pharmacol. 2002, 135, 356-362.). Serum levels of CGRP increased during migraine (Goadsby PJ, et al. Ann Neurol 1990; 28: 183-7), and treatment with anti-migraine drugs reduced headache to normal CGRP levels Return (Gallai V. et al. Cephalalgia 1995; 15: 384-90). Migraine patients show elevated baseline CGRP levels compared to control patients (Ashina M, et al., Pain 2000, 86 (1-2): 133-8.2000). Intravenous CGRP inhalation sustains the headache in migraine patients (Lassen LH, et al. Cephalalgia 2002 Feb; 22 (1): 54-61). Preclinical studies in dogs and rats have reported that systemic CGRP blockade by the peptide antagonist CGRP (8-37) does not alter resting systemic hemodynamics or local blood flow (Shen, YT. Et al, J Pharmacol Exp Ther 2001, 298, 551-8). Thus, CGRP receptor antagonists provide a novel treatment for migraine that avoids the likelihood of active cardiovascular vasoconstriction associated with non-selective 5-HT1B / 1D agonists, “triptans” (eg, sumatriptan). It can be shown.

  CGRP antagonists have shown efficacy in human clinical trials. Davis CD, Xu C. Curr Top Med Chem. 2008 8 (16): 1468-79; Benemei S, Nicoletti P, Capone JG, Geppetti P. Curr Opin Pharmacol. 2009 9 (1): 9-14. Epub 2009 Jan 20; Ho TW, Ferrari MD, Dodick DW, Galet V, Kost J, Fan X, Leibensperger H, Froman S, Assaid C, Lines C, Koppen H, Winner PK. Lancet. 2008 372: 2115. Epub 2008 Nov 25; See Ho TW, Mannix LK, Fan X, Assaid C, Furtek C, Jones CJ, Lines CR, Rapoport AM; Neurology 2008 70: 1304. Epub 2007 Oct 3.

Pharmaceutical compositions and methods of treatment The hemisulfate of compound (I) inhibits the CGRP receptor. Thus, the hemisulfate salt of Compound (I) is useful for treating diseases or disorders associated with abnormal CGRP levels or where modulation of CGRP levels may have therapeutic benefit.

  Accordingly, another aspect of the invention is a pharmaceutical composition comprising a hemisulfate salt of Compound (I) together with a pharmaceutically acceptable adjuvant, carrier or diluent.

  Certain embodiments provide a pharmaceutical composition comprising a hemisulfate salt of Compound (I) sesquihydrate together with a pharmaceutically acceptable adjuvant, carrier or diluent.

  Certain embodiments provide a pharmaceutical composition comprising a crystalline form of the hemisulfate salt of Compound (I) together with a pharmaceutically acceptable adjuvant, carrier, or diluent.

  Certain embodiments provide a pharmaceutical composition comprising a crystalline form of the hemisulfate salt of Compound (I) sesquihydrate, a pharmaceutically acceptable adjuvant, carrier, or diluent.

  Certain embodiments provide a pharmaceutical composition comprising hemisulfate Form H1.5-1 of Compound (I) together with a pharmaceutically acceptable adjuvant, carrier, or diluent.

  The compound is generally applied as a pharmaceutical composition comprised of a therapeutically effective amount of the hemisulfate salt of Compound (I) and a pharmaceutically acceptable carrier, and may contain conventional excipients. Good. A therapeutically effective amount is that amount needed to provide a meaningful patient benefit as determined by a medical practitioner in the art. Pharmaceutically acceptable carriers are generally known carriers that have an acceptable safety profile. Compositions include all common solid and liquid forms, including capsules, tablets, troches and powders, and liquid suspensions, syrups, lyxyls and solutions. Solid compositions can be formed in timed or sustained release formulations. The compositions are prepared using common formulation techniques, and conventional excipients (such as binders and wetting agents) and vehicles (such as water and alcohols).

  Solid compositions are usually formulated in dosage units that provide about 1 to about 1000 mg of active ingredient per dose. Examples of solid dosage units are 0.1 mg, 1 mg, 10 mg, 100 mg, 500 mg, and 1000 mg. Liquid compositions are generally in unit dosage ranges of 1-100 mg / mL. Examples of liquid dose units are 0.1 mg / mL, 1 mg / mL, 10 mg / mL, 25 mg / mL, 50 mg / mL, and 100 mg / mL.

  In certain embodiments, the oral dosage form provides 70-750 mg of Compound (I) as the hemisulfate salt of Compound (I). In this embodiment, oral dosage forms having 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 500 mg, and 750 mg of Compound (I) as the hemisulfate salt of Compound (I) are included.

  In certain embodiments, the oral dosage form provides 70-750 mg of Compound (I) as the hemisulfate form of Compound (I) H1.5-1. In this embodiment, the hemisulfate Form H1.5-1 of Compound (I) is oral having 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 500 mg, and 750 mg of Compound (I) Dosage forms are included.

  In certain embodiments, the hemisulfate salt of Compound (I) is administered once daily. Suitable doses include 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 500 mg, and 750 mg of Compound (I) as Compound (I) hemisulfate Form H1.5-1. .

  In certain embodiments, the hemisulfate salt of Compound (I) is administered twice daily. Suitable doses include 70 mg, 80 mg, 90 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 500 mg, and 750 mg of Compound (I) as Compound (I) hemisulfate Form H1.5-1. .

  The present invention encompasses all conventional modes of administration, including oral, parenteral, intranasal, sublingual, and transdermal modes. Typically, the once daily dose is 0.01-100 mg / kg body weight per day. In general, more compound is required orally and less amount of compound is required parenterally. However, the specific dosing schedule should be determined by a physician according to sound medical judgment.

  Inhibitors at the receptor level for CGRP are envisioned to be useful in pathophysiological conditions that result in excessive CGRP receptor activation. These include neurogenic vasodilation, neurogenic inflammation, migraine, cluster headache and other headaches, burns, circulatory shock, menopausal flush, and asthma. Activation of the CGRP receptor has been implicated in the pathogenesis of migraine (Edvinsson L. CNS Drugs 2001, 15 (10), 745-53; Williamson, DJ Microsc. Res. Tech. 2001, 53, 167- 178 .; Grant, AD Brit. J. Pharmacol. 2002, 135, 356-362.). Serum levels of CGRP increased during migraine (Goadsby PJ et al. Ann. Neurol. 1990, 28, 183-7), and treatment with anti-migraine drugs reduced headache and normalized CGRP levels. (Gallai V. et al. Cephalalgia 1995, 15, 384-90). Migraine patients show elevated baseline CGRP levels compared to control patients (Ashina M. et al., Pain 2000, 86 (1-2), 133-8). Intravenous CGRP inhalation sustains headaches in migraine patients (Lassen L.H. et al. Cephalalgia. 2002, 22 (1), 54-61). Preclinical studies in dogs and rats have reported that systemic CGRP blockade by the peptide antagonist CGRP (8-37) does not alter resting systemic hemodynamics or local blood flow (Shen, YT. Et al. J Pharmacol. Exp. Ther. 2001, 298, 551-8). Thus, CGRP receptor antagonists are novel treatments for migraine that avoid the likelihood of active cardiovascular vasoconstriction associated with non-selective 5-HT1B / 1D agonist “triptans” (eg, sumatriptan). Can be shown.

  In another aspect, the present invention is a method for inhibiting a CGRP receptor, comprising contacting the CGRP receptor with a hemisulfate salt of compound (I).

  An embodiment provides a method for inhibiting a CGRP receptor, comprising contacting the CGRP receptor with a hemisulfate salt of Compound (I) sesquihydrate.

  One embodiment provides a method for inhibiting a CGRP receptor, comprising contacting the CGRP receptor with crystalline hemisulfate of Compound (I) sesquihydrate.

  One embodiment provides a method of inhibiting a CGRP receptor, comprising contacting the CGRP receptor with the hemisulfate form H1.5-1 of compound (I).

  In another aspect, the present invention is a method for treating a disease associated with abnormal levels of CGRP, comprising administering to a patient a therapeutically effective amount of a hemisulfate salt of Compound (I).

  In another aspect, the invention is the use of a hemisulfate salt of Compound (I) in the manufacture of a therapeutic agent for a disease associated with abnormal levels of CGRP.

  In another aspect, the invention is a method for treating migraine or headache.

  Other aspects of the invention include inflammation (especially neuropathic inflammation), pain, burns, circulatory shock, diabetes, Raynaud's syndrome, peripheral arterial failure, subarachnoid / intracranial hemorrhage, tumor growth, flushing associated with menopause and others Wherein the treatment is achieved by antagonism of the CGRP receptor by administration of a pharmaceutical composition comprising a hemisulfate salt of compound (I) as defined herein. Can do.

  Another aspect of the invention includes (a) immunomodulation in the intestinal mucosa, (b) protective effect against cardiac anaphylactic injury, (c) activity or inhibition of interleukin-1b (IL-1b) stimulation of bone resorption, ( d) modulation of NK1 receptor expression in spinal neurons and (e) methods selected from the group consisting of airway inflammatory diseases and chronic obstructive pulmonary diseases (including asthma). (a) Calcitonin Receptor-Like Receptor Is Expressed on Gastrointestinal Immune Cells.Hagner, Stefanie; Knauer, Jens; Haberberger, Rainer; Goeke, Burkhard; Voigt, Karlheinz; McGregor, Gerard Patrick. Digestion (2002), 66 (4), 197-203; (b) Protective effects of calcitonin gene-related peptide-mediated evodiamine on guinea-pig cardiac anaphylaxis.Rang, Wei-Qing; Du, Yan-Hua; Hu, Chang-Ping; Ye, Feng; Tan, Gui-Shan; Deng, Han-Wu; Li, Yuan-Jian.School of Pharmaceutical Sciences, Department of Pharmacology, Central South University, Xiang-Ya Road 88, Changsha, Hunan, Naunyn -Schmiedeberg's Archives of Pharmacology (2003), 367 (3), 306-311; (c) The experimental study on the effect calcitonin gene-related peptide on bone resorption mediated by interleukin-1. Lian, Kai; Du, Jingyuan; Rao , Zhenyu; Luo, Huaican.Department of Orthopedics, Xiehe Hospital, Tongji Medical College, Huazhong University of Science and Technology, W uhan, Peop. Rep. China.Journal of Tongji Medical University (2001), 21 (4), 304-307, (d) Calcitonin gene-related Peptide regulates expression of neurokinin1 receptors by rat spinal neurons.Seybold VS, McCarson KE, Mermelstein PG, Groth RD, Abrahams LG. J. Neurosci. 2003 23 (5): 1816-1824.Department of Neuroscience, University of Minnesota, Minneapolis, Minnesota 55455, and Department of Pharmacology, Toxicology, and Therapeutics, University of Kansas Medical Center, Kansas City, Kansas 66160 (e) Attenuation of antigen-induced airway hyperresponsiveness in CGRP-deficient mice.Aoki-Nagase, Tomoko; Nagase, Takahide; Oh-Hashi, Yoshio; Shindo, Takayuki; Kurihara, Yukiko; Yamaguchi, Yasuhiro ; Yamamoto, Hiroshi; Tomita, Tetsuji; Ohga, Eijiro; Nagai, Ryozo; Kurihara, Hiroki; Ouchi, Yasuyoshi. Department of Geriatric Medicine, Graduate School of Medicine, University of Tokyo, Tokyo, Japan. American Journal of Physiology (2002) , 283 (5, Pt. 1), L963-L970; (f) Calcitonin gene-related pep Springer, Jochen; Geppetti, Pierangelo; Fischer, Axel; Groneberg, David A. Charite Campus-Virchow, Department of Pediatric Pneumology and Immunology, Division of Allergy Research, Humboldt-University Berlin, Berlin, Germany. Pulmonary Pharmacology & Therapeutics (2003), 16 (3), 121-130; and (g) Pharmacological targets for the inhibition of neurogenic inflammation. Helyes, Zsuzsanna; Pinter, Erika; Nemeth, Jozsef; Szolcsanyi, Janos. Department of Pharmacology and Pharmacotherapy, See Faculty of Medicine, University of Pecs, Pecs, Hung. Current Medicinal Chemistry: Anti-Inflammatory & Anti-Allergy Agents (2003), 2 (2), 191-218.

  In another aspect, the present invention is selected from the group consisting of compound (I) hemisulfate and a COX2 inhibitor, NSAIDS, aspirin, acetaminophen, triptan, ergotamine and caffeine for the treatment of migraine And a method of treatment using a combination with one or more other drugs.

  “Migraine”, “headache” and related terms are as understood by healthcare professionals. Migraine includes all classes of migraine, including typical, common, cluster, electric shock, hemiplegic, ocular palsy, and ocular.

  A “therapeutically effective amount”, when administered alone or in combination with further therapeutic agents, prevents and inhibits the disease and / or illness and / or progression of the disease and / or illness, and / or Means an amount that is effective to reduce.

  “Patient” means a person who can benefit from treatment as determined by a healthcare professional.

Examples The invention is further defined in the following examples. It should be understood that the examples are described for illustrative purposes only. From the above description and examples, those skilled in the art can ascertain the essential features of the present invention without departing from the spirit and scope thereof, and various modifications and variations to use the present invention in various applications and conditions. Modifications can be made. Accordingly, the invention is not limited by the exemplary embodiments described below, but rather is defined by the appended claims.

Abbreviations DMSO dimethyl sulfoxide EDTA ethylenediaminetetraacetic acid eq equivalent ESI electrospray ionization mass spectrometry g gram h time L liter LCMS liquid chromatography mass spectrometry M mole mg milligram min minute mL milliliter (s)
mmol mmol MS mass spectrometry N normal NaHMDS sodium bis (trimethylsilyl) amide NCS N-chlorosuccinimide NMR nuclear magnetic resonance spectroscopy RT retention time ssNMR solid nuclear magnetic resonance TBAF tetrabutylammonium fluoride TFA trifluoroacetic acid THF tetrahydrofuran TIPSO triisopropylsilyl Oxy TMS Tetramethylsilane μL Microliter ° C Celsius

Proton magnetic resonance ( ¹ H NMR) spectra were recorded on a Bruker AC300 or AC500. All spectra are determined in the indicated solvent, chemical shifts are described in δ units low magnetic field from the internal standard tetramethylsilane (TMS), and interproton coupling constants are described in hertz (Hz). . The splitting pattern is expressed as follows: s, singlet; d, doublet; t, triplet; q, quartet; m, multiplet; br, broad peak. Low resolution mass spectra (MS) and apparent molecular weights (MH ⁺ ) or (MH) ⁺ were examined on a Micromass platform. Elemental analysis is stated as weight percent. The product starts with a column YMC S5 ODS (30 × 100 mm) with a solvent composition of 40% methanol-60% water-0.1% TFA with a flow rate of 40.0 mL / min and a gradient of 8.0 minutes, Purification by preparative HPLC using solvent composition up to 95% methanol-5% water-0.1% TFA. The product starts with an XTERA column (3.0 × 50 mm S7) starting with solvent A (10% methanol-90% water-0.1% trifluoroacetic acid (TFA)) over a 2 minute gradient time, followed by solvent B ( 10% water-90% methanol-0.1% TFA) and analyzed by HPLC. The flow rate was 5 mL / min and the product retention time (Rf) was measured at 220 nm wavelength.

（中間体１）
２５０ｍＬの丸底フラスコにおいて、テトラヒドロフラン（５ｍＬ）中に（９Ｒ）−６−（２，３−ジフルオロフェニル）−９−（トリイソプロピルシリルオキシ）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−５−オン（０．２１８ｇ，０．４９ｍｍｏｌ）を溶解させて無色の溶液を得た。窒素下で−１５℃（氷−メタノール槽）に冷却し、ＴＢＡＦ（０．４９０ｍＬ，０．４９０ｍｍｏｌ）を加え、得られた明るい黄色の溶液を−１５℃で１時間攪拌した。これを炭酸水素ナトリウム溶液でクエンチし、酢酸エチルで希釈した。層を分離し、水層を酢酸エチルで抽出した。有機層を合わせて、食塩水で洗浄し、乾燥させ、濃縮して、黄褐色の油状物を得た。１００％以下の酢酸エチル／ヘキサンを用いてフラッシュカラムクロマトグラフィー（２５ｇシリカゲルカラム）により、所望生成物を得た（１１２ｍｇ，６２％）。¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.53 (dd, J=4.91, 1.64 Hz, 1 H) 7.85 (dd, J=7.68, 1.64 Hz, 1 H) 7.34 (dd, J=7.68, 4.91 Hz, 1 H) 7.00-7.16 (m, 3 H) 5.32 (s, 1 H) 4.94-5.04 (m, 1 H) 4.48 (dd, J=11.83, 3.02 Hz, 1 H) 2.14-2.48 (m, 4 H); 19F NMR (376 MHz, クロロホルム-d) δ ppm -138.24--138.07 (m, 1 F) -140.70--140.50 (m, 1 F). Intermediate 1
(6S, 9R) -6- (2,3-Difluorophenyl) -9-hydroxy-6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-5-one

(Intermediate 1)
In a 250 mL round bottom flask, (9R) -6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H-cyclohepta in tetrahydrofuran (5 mL). [B] Pyridin-5-one (0.218 g, 0.49 mmol) was dissolved to obtain a colorless solution. Cool to −15 ° C. (ice-methanol bath) under nitrogen, add TBAF (0.490 mL, 0.490 mmol) and stir the resulting bright yellow solution at −15 ° C. for 1 hour. This was quenched with sodium bicarbonate solution and diluted with ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic layers were combined, washed with brine, dried and concentrated to give a tan oil. Flash column chromatography (25 g silica gel column) using up to 100% ethyl acetate / hexanes gave the desired product (112 mg, 62%). ¹ H NMR (400 MHz, chloroform-d) δ ppm 8.53 (dd, J = 4.91, 1.64 Hz, 1 H) 7.85 (dd, J = 7.68, 1.64 Hz, 1 H) 7.34 (dd, J = 7.68, 4.91 Hz, 1 H) 7.00-7.16 (m, 3 H) 5.32 (s, 1 H) 4.94-5.04 (m, 1 H) 4.48 (dd, J = 11.83, 3.02 Hz, 1 H) 2.14-2.48 (m, 4 H); 19F NMR (376 MHz, chloroform-d) δ ppm -138.24--138.07 (m, 1 F) -140.70--140.50 (m, 1 F).

（中間体２）
水素化ホウ素リチウム（０．９８２ｇ，４５．１ｍｍｏｌ）を、Ｎ_２下にて０℃で（６Ｓ，９Ｒ）−６−（２，３−ジフルオロフェニル）−９−（トリイソプロピルシリルオキシ）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−５−オン（５．０２２４ｇ，１１．２７ｍｍｏｌ）のシクロペンチルメチルエーテル（３０ｍＬ）溶液に加えた。反応混合液を０℃で２時間攪拌し、続いて室温でさらに４時間攪拌した。該反応物を、メタノールを加えてクエンチした。反応混合液を０．５時間攪拌した。該溶媒を減圧下でほぼ留去し、粗物質を酢酸エチル中に入れ、水で３回洗浄した。０〜１０％のヘキサン中の酢酸エチルによるフラッシュカラムで所望生成物を得た（３．２８ｇ，６５％）。 Intermediate 2
(5S, 6S, 9R) -6- (2,3-Difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-5-ol.

(Intermediate 2)
Lithium borohydride (0.982 g, 45.1 mmol) was added (6S, 9R) -6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6 at 0 ° C. under N _2. , 7,8,9-Tetrahydro-5H-cyclohepta [b] pyridin-5-one (5.0224 g, 11.27 mmol) in cyclopentyl methyl ether (30 mL). The reaction mixture was stirred at 0 ° C. for 2 hours, followed by further stirring at room temperature for 4 hours. The reaction was quenched by adding methanol. The reaction mixture was stirred for 0.5 hour. The solvent was nearly distilled off under reduced pressure and the crude material was taken up in ethyl acetate and washed three times with water. The desired product was obtained on a flash column with 0-10% ethyl acetate in hexane (3.28 g, 65%).

（中間体３）
オーブンで乾燥させた２５０ｍＬの丸底フラスコにおいて、ＮＣＳ（０．７５１ｇ，５．６２ｍｍｏｌ）をテトラヒドロフラン（１５ｍＬ）中で懸濁させた。トリフェニルホスフィン（１．４７５ｇ，５．６２ｍｍｏｌ）を加えた。窒素下で５分間攪拌し、（５Ｓ，６Ｓ，９Ｒ）−６−（２，３−ジフルオロフェニル）−９−（トリイソプロピルシリルオキシ）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−５−オール（１．００７ｇ，２．２５０ｍｍｏｌ）を灰色の懸濁液に一度に加えた。生じた赤みがかった懸濁液を室温で攪拌した。該固形物が徐々に溶解して黄褐色の溶液を得た。５時間後、ＬＣＭＳにより変換が完了したことを示された。テトラヒドロフランを減圧中で留去し、残った赤色の油状物を６０％以下の酢酸エチル／ヘキサンによるＩＳＣＯ（２４０ｇシリカカラム）によって直接精製した。純粋な酢酸エチルで溶出した非極性成分および生成物を、塩化メチレン中の１０％ メタノール（２．０Ｍ ＮＨ_４ＯＨ）で溶出した。生成フラクションを合わせて、５０％ 酢酸エチル／ヘキサンでＦＣＣによって再精製して、無色の油状物として所望生成物を得た（８６９ｍｇ，８３％）。MS(ESI)[M+H⁺] = 466.22; ¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.55 (d, J=3.53 Hz, 1 H) 7.63 (br. s., 1 H) 7.20 (dd, J=7.68, 4.91 Hz, 1 H) 7.01-7.15 (m, 1 H) 6.90-7.01 (m, 1 H) 6.66-6.90 (m, 1 H) 5.55-5.85 (m, 1 H) 5.40-5.56 (m, 1 H) 3.96-4.33 (m, 1 H) 2.33 (br. s., 3 H) 2.09-2.20 (m, 1 H) 1.14-1.23 (m, 3 H) 1.04-1.14 (m, 9 H) 1.01 (d, J=7.30 Hz, 9 H). Intermediate 3
(5R, 6S, 9R) -5-chloro-6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridine

(Intermediate 3)
NCS (0.751 g, 5.62 mmol) was suspended in tetrahydrofuran (15 mL) in a 250 mL round bottom flask dried in oven. Triphenylphosphine (1.475 g, 5.62 mmol) was added. The mixture was stirred for 5 minutes under nitrogen, and (5S, 6S, 9R) -6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H-cyclohepta [ b] Pyridin-5-ol (1.007 g, 2.250 mmol) was added in one portion to the gray suspension. The resulting reddish suspension was stirred at room temperature. The solid gradually dissolved to give a tan solution. After 5 hours, LCMS indicated that the conversion was complete. Tetrahydrofuran was removed in vacuo and the remaining red oil was purified directly by ISCO (240 g silica column) with less than 60% ethyl acetate / hexane. Nonpolar components and product eluted with pure ethyl acetate were eluted with 10% methanol in methylene chloride (2.0 M NH ₄ OH). The product fractions were combined and repurified by FCC with 50% ethyl acetate / hexanes to give the desired product as a colorless oil (869 mg, 83%). MS (ESI) [M + H ⁺ ] = 466.22; ¹ H NMR (400 MHz, chloroform-d) δ ppm 8.55 (d, J = 3.53 Hz, 1 H) 7.63 (br. S., 1 H) 7.20 ( dd, J = 7.68, 4.91 Hz, 1 H) 7.01-7.15 (m, 1 H) 6.90-7.01 (m, 1 H) 6.66-6.90 (m, 1 H) 5.55-5.85 (m, 1 H) 5.40- 5.56 (m, 1 H) 3.96-4.33 (m, 1 H) 2.33 (br. S., 3 H) 2.09-2.20 (m, 1 H) 1.14-1.23 (m, 3 H) 1.04-1.14 (m, 9 H) 1.01 (d, J = 7.30 Hz, 9 H).

（中間体４）
１００ｍＬの丸底フラスコにおいて、（５Ｒ，６Ｓ，９Ｒ）−５−クロロ−６−（２，３−ジフルオロフェニル）−９−（トリイソプロピルシリルオキシ）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン（５６６ｍｇ，１．２１４ｍｍｏｌ）をジメチルホルムアミド（５ｍＬ）中に溶解させて、無色の溶液を得た。アジ化ナトリウム（４７４ｍｇ，７．２９ｍｍｏｌ）を加え、該混合物を窒素下にて室温で２．５時間攪拌した。ＬＣＭＳにより一部のみの反応が示された。該混合物を５０℃で終夜加熱した。１５時間後、ＬＣＭＳによりいくらかの除去生成物と共に完全な変換が示された。該混合物を水および酢酸エチルで希釈した。該層を分離した。有機層を食塩水で洗浄し、乾燥させ、濃縮して、無色の油状物を得た。粗生成物をさらに精製し、特徴付けをすることなく、次の反応に用いた。小規模の精製により分析試料を得た：MS(ESI)[M+H⁺] = 473.27; ¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.52-8.63 (m, 1 H) 7.75 (d, J=7.81 Hz, 1 H) 7.23-7.36 (m, 1 H) 6.95-7.17 (m, 2 H) 6.89 (br. s., 1 H) 5.28 (d, J=4.03 Hz, 1 H) 4.90 (d, J=9.07 Hz, 1 H) 3.79 (t, J=9.44 Hz, 1 H) 1.86-2.23 (m, 4 H) 1.16-1.30 (m, 3 H) 0.98-1.15 (m, 18 H); 19F NMR (376 MHz, クロロホルム-d) δ ppm -137.68--137.36 (m, 1 F) -141.78--141.54 (m, 1 F). Intermediate 4
(5S, 6S, 9R) -5-Azido-6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridine

(Intermediate 4)
In a 100 mL round bottom flask, (5R, 6S, 9R) -5-chloro-6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H -Cyclohepta [b] pyridine (566 mg, 1.214 mmol) was dissolved in dimethylformamide (5 mL) to give a colorless solution. Sodium azide (474 mg, 7.29 mmol) was added and the mixture was stirred at room temperature for 2.5 hours under nitrogen. LCMS showed only a partial reaction. The mixture was heated at 50 ° C. overnight. After 15 hours, LCMS showed complete conversion with some removed product. The mixture was diluted with water and ethyl acetate. The layers were separated. The organic layer was washed with brine, dried and concentrated to give a colorless oil. The crude product was further purified and used in the next reaction without characterization. An analytical sample was obtained by small-scale purification: MS (ESI) [M + H ⁺ ] = 473.27; ¹ H NMR (400 MHz, chloroform-d) δ ppm 8.52-8.63 (m, 1 H) 7.75 (d, J = 7.81 Hz, 1 H) 7.23-7.36 (m, 1 H) 6.95-7.17 (m, 2 H) 6.89 (br. S., 1 H) 5.28 (d, J = 4.03 Hz, 1 H) 4.90 ( d, J = 9.07 Hz, 1 H) 3.79 (t, J = 9.44 Hz, 1 H) 1.86-2.23 (m, 4 H) 1.16-1.30 (m, 3 H) 0.98-1.15 (m, 18 H); 19F NMR (376 MHz, chloroform-d) δ ppm -137.68--137.36 (m, 1 F) -141.78--141.54 (m, 1 F).

（中間体５）
１００ｍＬの丸底フラスコにおいて、（５Ｓ，６Ｓ，９Ｒ）−５−アジド−６−（２，３−ジフルオロフェニル）−９−（トリイソプロピルシリルオキシ）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン（０．７３２ｇ，１．５４９ｍｍｏｌ）（粗製）をテトラヒドロフラン（８ｍＬ）中に溶解させて、無色の溶液を得た。ＴＢＡＦ（１．８５９ｍＬ，１．８５９ｍｍｏｌ）を加え、得られた淡黄色の溶液を室温で１．５時間攪拌した。ＬＣＭＳにより変換が完了したことが示された。テトラヒドロフランを留去し、残渣を水および酢酸エチルで希釈した。該層を分離した。有機層を食塩水で洗浄し、乾燥させ、濃縮して、淡黄色の油状物を得た。６０％ 酢酸エチル／ヘキサンのＦＣＣによる精製により、所望生成物（粗製重量：４８０ｍｇ）を無色の油状物として得た。小規模精製により、分析試料を得た：MS(ESI)[M+H⁺] = 317.22; ¹H NMR (400 MHz, クロロホルム-d) δ ppm 8.51 (dd, J=4.91, 1.38 Hz, 1 H) 7.99 (d, J=7.30 Hz, 1 H) 7.35 (dd, J=7.81, 5.04 Hz, 1 H) 7.06-7.20 (m, 2 H) 6.94-7.05 (m, 1 H) 5.91 (br. s., 1 H) 5.03 (d, J=10.32 Hz, 1 H) 4.92 (dd, J=11.21, 2.39 Hz, 1 H) 2.84-3.02 (m, 1 H) 2.37-2.49 (m, 1 H) 2.25-2.36 (m, 1 H) 2.07-2.17 (m, J=14.38, 4.94, 3.05, 3.05 Hz, 1 H) 1.40-1.64 (m, 1 H); ¹³C NMR (101 MHz, クロロホルム-d) δ ppm 158.48 (s, 1 C) 152.19-149.87 (dd, J=13.10 and 221Hz, 1 C) 149.72-147.42 (dd, J=13.87および219 Hz, 1 C) 146.16 (s, 3 C) 133.67 (s, 2 C) 133.23 (s, 1 C) 132.66 (d, J=10.79 Hz, 1 C) 124.43 (dd, J=6.94, 3.85 Hz, 2 C) 123.84 (br. s., 1 C) 122.89 (s, 2 C) 115.98 (d, J=17.73 Hz, 2 C) 70.94 (s, 3 C) 65.67 (s, 1 C) 45.43 (br. s., 1 C) 35.71 (s, 3 C) 33.45 (s, 2 C); 19F NMR (376 MHz, クロロホルム-d) δ ppm -137.55--137.20 (m, 1 F) -142.28--141.89 (m, 1 F). Intermediate 5
(5S, 6S, 9R) -5-Azido-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-ol

(Intermediate 5)
In a 100 mL round bottom flask, (5S, 6S, 9R) -5-azido-6- (2,3-difluorophenyl) -9- (triisopropylsilyloxy) -6,7,8,9-tetrahydro-5H -Cyclohepta [b] pyridine (0.732 g, 1.549 mmol) (crude) was dissolved in tetrahydrofuran (8 mL) to give a colorless solution. TBAF (1.859 mL, 1.859 mmol) was added and the resulting pale yellow solution was stirred at room temperature for 1.5 hours. LCMS showed that the conversion was complete. Tetrahydrofuran was distilled off and the residue was diluted with water and ethyl acetate. The layers were separated. The organic layer was washed with brine, dried and concentrated to give a pale yellow oil. Purification by FCC with 60% ethyl acetate / hexanes gave the desired product (crude weight: 480 mg) as a colorless oil. An analytical sample was obtained by small-scale purification: MS (ESI) [M + H ⁺ ] = 317.22; ¹ H NMR (400 MHz, chloroform-d) δ ppm 8.51 (dd, J = 4.91, 1.38 Hz, 1 H ) 7.99 (d, J = 7.30 Hz, 1 H) 7.35 (dd, J = 7.81, 5.04 Hz, 1 H) 7.06-7.20 (m, 2 H) 6.94-7.05 (m, 1 H) 5.91 (br. S ., 1 H) 5.03 (d, J = 10.32 Hz, 1 H) 4.92 (dd, J = 11.21, 2.39 Hz, 1 H) 2.84-3.02 (m, 1 H) 2.37-2.49 (m, 1 H) 2.25 -2.36 (m, 1 H) 2.07-2.17 (m, J = 14.38, 4.94, 3.05, 3.05 Hz, 1 H) 1.40-1.64 (m, 1 H); ¹³ C NMR (101 MHz, chloroform-d) δ ppm 158.48 (s, 1 C) 152.19-149.87 (dd, J = 13.10 and 221Hz, 1 C) 149.72-147.42 (dd, J = 13.87 and 219 Hz, 1 C) 146.16 (s, 3 C) 133.67 (s, 2 C) 133.23 (s, 1 C) 132.66 (d, J = 10.79 Hz, 1 C) 124.43 (dd, J = 6.94, 3.85 Hz, 2 C) 123.84 (br. S., 1 C) 122.89 (s, 2 C) 115.98 (d, J = 17.73 Hz, 2 C) 70.94 (s, 3 C) 65.67 (s, 1 C) 45.43 (br. S., 1 C) 35.71 (s, 3 C) 33.45 (s, 2C); 19F NMR (376 MHz, chloroform-d) δ ppm -137.55--137.20 (m, 1 F) -142.28--141.89 (m, 1 F).

（中間体６）
１００ｍＬの丸底フラスコにおいて、（５Ｓ，６Ｓ，９Ｒ）−５−アジド−６−（２，３−ジフルオロフェニル）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−９−オール（０．４９０ｇ，１．５４９ｍｍｏｌ）（乾燥ベンゼンと共沸）および４−ニトロフェニル ４−（２−オキソ−２，３−ジヒドロ−１Ｈ−イミダゾ［４，５−ｂ］ピリジン−１−イル）ピペリジン−１−カルボキシレート（０．７１３ｇ，１．８５９ｍｍｏｌ）を、ジメチルホルムアミド（８ｍＬ）中に溶解させて、窒素下で淡黄色の懸濁液を得た。−１５℃（氷−メタノール槽）に冷却し、ＮａＨＭＤＳ（４．１８ｍＬ，４．１８ｍｍｏｌ）を滴下して加えた。生じた黄褐色の溶液を、窒素下にて−１０℃〜０℃で２時間、続いて室温で２時間攪拌した。ＬＣＭＳにより完全な変換が示された。該反応を炭酸水素ナトリウム溶液でクエンチした。該混合物を酢酸エチルで希釈した。層を分離し、水層を酢酸エチルで抽出した。有機層を合わせて、水、食塩水で洗浄し、硫酸ナトリウムで乾燥させ、濃縮して、黄褐色の油状物を得た。８％ メタノール／塩化メチレンでＦＣＣによる精製により、所望生成物（主要なピーク，６３２ｍｇ，３段階で７３％）を淡黄色の泡状物として得た。MS(ESI)[M+H⁺] = 561.27; ¹H NMR (400 MHz, クロロホルム-d) δ ppm 11.50 (br. s., 1 H) 8.58 (d, J=3.78 Hz, 1 H) 8.11 (d, J=5.04 Hz, 1 H) 7.91 (d, J=7.30 Hz, 1 H) 7.33 (br. s., 2 H) 7.07-7.19 (m, 2 H) 6.92-7.06 (m, 2 H) 6.10 (d, J=9.32 Hz, 1 H) 5.23 (d, J=10.07 Hz, 1 H) 4.26-4.84 (m, 3 H) 2.46-3.34 (m, 4 H) 2.20-2.43 (m, 3 H) 2.01-2.13 (m, 1 H) 1.94 (d, J=12.34 Hz, 3 H); 19F NMR (376 MHz, クロロホルム-d) δ ppm -137.30--137.01 (m, 1 F) -142.32--142.03 (m, 1 F). Intermediate 6
(5S, 6S, 9R) -5-Azido-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl 4- (2-oxo -2,3-Dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate

(Intermediate 6)
In a 100 mL round bottom flask, (5S, 6S, 9R) -5-azido-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridine-9- All (0.490 g, 1.549 mmol) (azeotrope with dry benzene) and 4-nitrophenyl 4- (2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl ) Piperidine-1-carboxylate (0.713 g, 1.859 mmol) was dissolved in dimethylformamide (8 mL) to give a pale yellow suspension under nitrogen. Cool to −15 ° C. (ice-methanol bath) and add NaHMDS (4.18 mL, 4.18 mmol) dropwise. The resulting tan solution was stirred under nitrogen at −10 ° C. to 0 ° C. for 2 hours followed by 2 hours at room temperature. LCMS showed complete conversion. The reaction was quenched with sodium bicarbonate solution. The mixture was diluted with ethyl acetate. The layers were separated and the aqueous layer was extracted with ethyl acetate. The organic layers were combined, washed with water, brine, dried over sodium sulfate and concentrated to give a tan oil. Purification by FCC with 8% methanol / methylene chloride gave the desired product (major peak, 632 mg, 73% over 3 steps) as a pale yellow foam. MS (ESI) [M + H ⁺ ] = 561.27; ¹ H NMR (400 MHz, chloroform-d) δ ppm 11.50 (br. S., 1 H) 8.58 (d, J = 3.78 Hz, 1 H) 8.11 ( d, J = 5.04 Hz, 1 H) 7.91 (d, J = 7.30 Hz, 1 H) 7.33 (br. s., 2 H) 7.07-7.19 (m, 2 H) 6.92-7.06 (m, 2 H) 6.10 (d, J = 9.32 Hz, 1 H) 5.23 (d, J = 10.07 Hz, 1 H) 4.26-4.84 (m, 3 H) 2.46-3.34 (m, 4 H) 2.20-2.43 (m, 3 H ) 2.01-2.13 (m, 1 H) 1.94 (d, J = 12.34 Hz, 3 H); 19F NMR (376 MHz, chloroform-d) δ ppm -137.30--137.01 (m, 1 F) -142.32-- 142.03 (m, 1 F).

（Ｉ）
１００ｍＬの丸底フラスコにおいて、（５Ｓ，６Ｓ，９Ｒ）−５−アジド−６−（２，３−ジフルオロフェニル）−６，７，８，９−テトラヒドロ−５Ｈ−シクロヘプタ［ｂ］ピリジン−９−イル ４−（２−オキソ−２，３−ジヒドロ−１Ｈ−イミダゾ［４，５−ｂ］ピリジン−１−イル）ピペリジン−１−カルボキシレート（６２０ｍｇ，１．１０６ｍｍｏｌ）（中間体６）を、テトラヒドロフラン（５ｍＬ）中に溶解させて、無色の溶液を得た。トリメチルホスフィン（３．３２ｍＬ，３．３２ｍｍｏｌ，トルエン中で１．０Ｍ）を加えた。該混合物を室温で攪拌した。２時間後、ＬＣＭＳにより出発物質が存在しないことが示された。水（０．０８０ｍＬ，４．４２ｍｍｏｌ）を加え、該混合物をさらに３時間攪拌した。ＬＣＭＳにより、所望生成物への完全な変換が示された。揮発性構成物質を減圧中で留去し、残渣を塩化メチレン中の１０％ メタノールでのＦＣＣで直接精製して、生成物（５１０ｍｇ，８５％）を白色の固形物として得た。MS(ESI)[M+H⁺] = 535.23; ¹H NMR (400 MHz, クロロホルム-d) δ ppm 10.39 (br. s., 1 H) 8.52 (d, J=3.78 Hz, 1 H) 8.09 (d, J=5.04 Hz, 2 H) 7.46 (br. s., 1 H) 7.26-7.38 (m, 1 H) 7.06-7.20 (m, 3 H) 6.94-7.05 (m, 1 H) 6.06-6.23 (m, 1 H) 4.31-4.78 (m, 4 H) 4.05 (spt, J=6.13 Hz, 1 H) 2.57-3.25 (m, 3 H) 2.17-2.38 (m, 3 H) 1.42-2.04 (m, 6 H); 19F NMR (376 MHz, クロロホルム-d) δ ppm -136.90 (br. s., 1 F) -142.48--142.21 (m, 1 F). Compound (I)
(5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl 4- (2-oxo -2,3-Dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate

(I)
In a 100 mL round bottom flask, (5S, 6S, 9R) -5-azido-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridine-9- Yl 4- (2-oxo-2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate (620 mg, 1.106 mmol) (intermediate 6) Dissolved in tetrahydrofuran (5 mL) to give a colorless solution. Trimethylphosphine (3.32 mL, 3.32 mmol, 1.0 M in toluene) was added. The mixture was stirred at room temperature. After 2 hours, LCMS showed no starting material present. Water (0.080 mL, 4.42 mmol) was added and the mixture was stirred for an additional 3 hours. LCMS showed complete conversion to the desired product. The volatile constituents were removed in vacuo and the residue was purified directly by FCC with 10% methanol in methylene chloride to give the product (510 mg, 85%) as a white solid. MS (ESI) [M + H ⁺ ] = 535.23; ¹ H NMR (400 MHz, chloroform-d) δ ppm 10.39 (br. S., 1 H) 8.52 (d, J = 3.78 Hz, 1 H) 8.09 ( d, J = 5.04 Hz, 2 H) 7.46 (br. s., 1 H) 7.26-7.38 (m, 1 H) 7.06-7.20 (m, 3 H) 6.94-7.05 (m, 1 H) 6.06-6.23 (m, 1 H) 4.31-4.78 (m, 4 H) 4.05 (spt, J = 6.13 Hz, 1 H) 2.57-3.25 (m, 3 H) 2.17-2.38 (m, 3 H) 1.42-2.04 (m , 6 H); 19F NMR (376 MHz, chloroform-d) δ ppm -136.90 (br. S., 1 F) -142.48--142.21 (m, 1 F).

High-throughput salt screening for crystalline salt of compound (I) High-throughput crystallization was used to screen for the formation of crystalline salt of compound (I). The screen tested acid type, acid level (equivalent), and / or type of crystallization solvent. Each plate contained 96 well plates (8 rows of 12 columns per plate).

A solution was prepared by dissolving 400 mg of compound (I) in a mixture of 36 mL THF and 4 mL H ₂ O. The solution (12.5 mL) was transferred to 24 vials. To each vial was added a 0.25M EtOH stock solution of the following acids:

  The contents of each vial were transferred to 12 crystallization wells and evaporated to dryness. By evaporation, each well was filled with 100 μl of solvent using a robotic liquid handler. The following crystallization solvents were tested: methyl isobutyl ketone (MIBK, ethyl acetate, toluene, THF, acetonitrile, acetone, isopropanol, ethanol, methanol, 1,2-dichloroethylene, isopropanol / water (50:50), and water. The plate was then sealed with a Teflon septum and subjected to a temperature cycle: the plate was kept at 50 ° C. for 10 hours, and then cooled to room temperature over 14 hours After the heating / cooling cycle, the well contents were duplicated. Characterized by refraction imaging, the recognized crystal hits were further characterized by PXRD analysis.

  Crystalline salt formation was not observed for Compound (I) in the presence of acetic acid, benzoic acid, benzenesulfonic acid, L-lactic acid, maleic acid, L-malic acid, phosphoric acid, and succinic acid. Crystalline salt formation was observed for compound (I) in the presence of citric acid, fumaric acid, hydrochloric acid, methanesulfonic acid, sulfuric acid, D-tartaric acid, and L-tartaric acid in at least one solvent. The characteristics of the crystalline salt of compound (I) were further characterized.

The results of salt screening and crystalline salt characterization are shown in Table 3.
Table 3

Example 1
(5S, 6S, 9R) -5-amino-6- (2,3-difluorophenyl) -6,7,8,9-tetrahydro-5H-cyclohepta [b] pyridin-9-yl 4- (2-oxo Preparation from -2,3-dihydro-1H-imidazo [4,5-b] pyridin-1-yl) piperidine-1-carboxylate, hemisulfate ethanol / water solution Compound (I) (1 g) was prepared at 70 ° C. Dissolved in 17 mL ethanol and water (3: 1) (solution A). Separately, 52 μL of 96% H ₂ SO ₄ (0.5 eq) was dissolved in 8 mL ethanol and water (3: 1) at room temperature (solution B). Next, 30 mg seed crystals were added to Solution A. Solution B was added to solution A seeded over 2 hours with a syringe pump. The resulting slurry was stirred at 70 ° C. for 1 hour and cooled to 20 ° C. over 90 minutes. The slurry was allowed to stir at room temperature overnight. The slurry was filtered. The wet cake was washed with 8 mL EtOH: water (3: 1) solution and dried in a vacuum oven at 30 ° C. overnight to give 1.01 g (88.4 mol%) of Example Compound 1 as crystals. Obtained as a solid. GADDS showed that the crystalline solid was Form H-1.5.

Preparation from tetrahydrofuran / water solution Compound (I) (1 g) was dissolved in 10 mL THF and water (4: 1) at 50 ° C. (solution A). Separately, 52 μL of 96% H ₂ SO ₄ (0.5 eq) was dissolved in 10 mL of THF at room temperature (Solution B). Next, 0.5 mL of Solution B was added to Solution A, followed by 20 mg of seed crystals. The solution turned into a thin slurry. The remaining amount of Solution B was slurried over 2 hours using a syringe pump. The slurry was stirred at 50 ° C. for 1 hour and subsequently cooled to 20 ° C. over 1 hour. The slurry was stirred overnight at room temperature. The slurry was filtered. The wet cake was washed with 8 mL THF: water = 3: 1 and dried in a vacuum oven at 30 ° C. overnight to give 1.06 g (92.8 mol%) of Example Compound 1 as a crystalline solid. Obtained. GADDS showed that the crystalline solid was Form H-1.5.

％ pot.＝効力％
％ total imp.＝合計不純物％
HIL／UV：高強度の光／紫外線 Stability Experiment The solid stability of Example Compound 1 was tested by exposing the sample to various temperature and relative humidity conditions for 1, 2 and 4 weeks. The efficacy% (% pot.) And total impurity% (% total imp.) Are shown in Table 4. The results show that the hemisulfate crystalline form H1.5-1 of compound (I) shows little increase in total impurity levels and / or no decrease in potency after 4 weeks of storage, It is shown to be stable under the storage conditions tested.
Table 4
Solid stability of hemisulfate form H1.5-1

% Pot. = Efficacy%
% Total imp. = Total impurity%
HIL / UV: High-intensity light / UV

  The moisture absorption isotherm for Example Compound 1 is shown in FIG. Example Compound 1 exhibits a hygroscopic weight gain of 0.8 wt% and 2.8 wt%, respectively, at a relative humidity between 25% and 75% and a relative humidity between 5% and 95%. These results indicated that the hemisulfate salt of Compound (I) has low or no hygroscopicity under the conditions tested.

Form H1.5-1
Table 5 is measured at about 25 ° C. for Example Compound 1 based on a high quality pattern collected on a diffractometer (CuKα) with a rotating capillary calibrated at 2θ using NIST and other suitable standards. The characteristic PXRD diffraction peak position (2θ ± 0.1 degrees) is shown.
Table 5
PXRD peak position (degree 2θ ± 0.1)

Table 6 shows the characteristic solid state NMR peak position δ (ppm) for Example Compound 1 standardized by TMS.
Table 6
ssNMR peak position-δ (ppm)

Other Crystal Forms of Example Compound 1 Form P22C: Form H1.5-1 was prepared by heating at 60 ° C. for 2 hours or 75 ° C. for 5 minutes. Water activity experiments between Form H1.5-1 and Form P22C showed that Form H1.5-1 is more stable at> 23% relative humidity.
Form P33: Form H1.5-1 converted to Form P33 between 50 ° C. and 75 ° C. in PXRD experiments at various temperatures. It was observed after heating Form H1.5-1 at 105 ° C. for 5 minutes or prepared by slurrying dry powder Form H1.5-1 in dry EtOH or IPAc. Elemental analysis indicated that Form P33 was hemisulfate monohydrate. Solid state NMR showed that Form P33 was a single phase. Water activity experiments between Form H1.5-1 and Form P33 showed that Form H1.5-1 is more stable at> 23% relative humidity.
Form P35: Prepared from a slurry of Form H1.5-1 in dry MeOH using Molecular Sieves (7% RH). Converted to Form P33 when dried at 60 ° C.

Stability in water slurry A water slurry of Example Compound 1 was prepared and stored at room temperature. After 2 days, there was little chemical degradation; there was no change in the PXRD pattern, indicating that crystalline form H1.5-1 was stable in the water slurry.

  There was little change in thermogravimetric scanning and differential scanning calorimetry and PXRD.

  The hemisulfate salt of compound (I) was compared with other salts of compound (I) and found to be particularly beneficial. The hemisulfate of compound (I) has the surprising effect of providing a salt that is physically stable and chemically stable compared to other salts of compound (I). Furthermore, hemisulfate has the surprising advantage provided by Form H1.5-1, which is a stable crystalline form. For example, the hemisulfate salt of Compound (I) is reproducibly prepared as a crystalline form, exhibits low hygroscopicity, and easily changes in crystalline form or hydration state in response to changes in relative humidity and / or temperature. There was no. In contrast, citrate, fumarate, hydrochloride, methanesulfonate, phosphate, and L-tartrate are hygroscopic at ambient temperature and relative humidity conditions, resulting in weight changes, water A change in sum state and / or a phase transition occurred. Crystalline salt formation is not seen for compound (I) in the presence of acetic acid, benzoic acid, benzenesulfonic acid, L-lactic acid, maleic acid, L-malic acid, and succinic acid in high throughput salt screening It was. Furthermore, the preparation of hemisulfate did not require the use of expensive materials such as D-tartaric acid.

Biological Methods In Vitro Pharmacological Tissue Culture SK-N-MC cells were prepared as monolayers in a medium consisting of L-glutamine (Invitrogen) supplemented with MEM containing Earl's salt and 10% fetal calf serum (Invitrogen). Grow at 37 ° C. in% CO ₂ .

Membrane preparation Crude membranes were prepared from SK-N-MC cells expressing CGRP receptors. The cells are rinsed twice with phosphate buffered saline (155 mM NaCl, 3.3 mM Na ₂ HPO ₄ , 1.1 mM KHPO ₄ , pH 7.4), low osmosis consisting of 10 mM Tris (pH 7.4) and 5 mM EDTA Incubated in pressure lysis buffer at 4 ° C. for 5-10 minutes. The cells were transferred from the plate to a polypropylene tube (16 × 100 mm) and homogenized using a polytron. The homogenized product was centrifuged at 32,000 × g for 30 minutes. The pellet was resuspended in cold hypotonic lysis buffer containing 0.1% mammalian protease inhibitor Cooktail (Sigma) and assayed for protein concentration. The SK-N-MC homogenized product was aliquoted and stored at -80 ° C.

Radioligand binding assay Compound (I) was solubilized and serially diluted with 100% DMSO. Aliquots from serial dilutions of compound were further diluted 25-fold with assay buffer (50 mM Tris-Cl pH 7.5, 5 mM MgCl ₂ , 0.005% Triton X-100) and (volume 50 μl) was 96 Transfer to well assay plate. [ ¹²⁵ I] -CGRP (GE Healthcare or Perkin-Elmer) was diluted to 72 pM with assay buffer and a volume of 50 μl was added to each well. The SK-N-MC membrane was thawed, diluted with assay buffer containing freshly prepared 0.1% mammalian protease inhibitor Cooktail (Sigma), and homogenized again. SK-N-MC homogenate (7 μg / well) was added in a volume of 100 μl. Subsequently, the assay plate was incubated for 2 hours at room temperature. Filter through a glass fiber filter (Whatman GF / B) presoaked in 0.5% PEI and immediately add an excess amount of cold wash buffer (50 mM Tris-Cl pH 7.5, 0.1% BSA). The assay was stopped by Non-specific binding was determined with 1 μM beta-CGRP (Bachem). Protein bound radioactivity was determined using a gamma or scintillation counter. The resulting data was analyzed using a four parameter competitive binding equation (XLfit v2.0) and IC ₅₀ was defined as the concentration of Compound (I) required to replace 50% of the radioligand binding. The final assay concentration of [ ¹²⁵ I] -CGRP was 18 pM. The average K _d for [ ¹²⁵ I] -CGRP is 25.4 pM. Compound (I) was evaluated in at least two separate experiments. In this experiment, the human CGRP receptor IC ₅₀ value of compound (I) was 0.04 nM.

In vivo pharmacokinetic experiments In vivo experiments were performed on the pharmacokinetics of the free base compound (I) in humans previously treated with 40 mg famotidine compared to untreated humans.

Compound (I) in human EDTA plasma was analyzed using liquid-liquid extraction with uHPLC-MS / MS detection on a Triple Quad 5500 mass spectrometer. This method utilized [ ¹³ C ₂ , D ₄ ] -compound (I) labeled with a stable isotope as an internal standard. 50 μL 1M NH ₄ OAc containing 100 ng / mL [ ¹³ C ₂ , D ₄ ] -Compound (I) and 4% acetate buffer solution in 50 μL MeOH: water (20/80) was added to each 0.100 mL. In addition to experimental samples, quality control (QC) samples, and calibration standards, the samples were extracted by stirring with 600 μL methyl tert-butyl ether (MTBE) for 15 minutes. 450 μL of the organic layer was removed and evaporated to dryness. The residue was reconstituted in 200 μL of reconstitution solution (30% acetonitrile in 10 mM NH ₄ OAc containing 0.01% acetic acid). All liquid transfer steps were performed using a Perkin Elmer JANUS Mini® liquid handler, except when adding an internal standard solution. A 10 μL aliquot of the extracted sample was injected into the uHPLC-MS / MS system. uHPLC was performed using a LEAP 4X Ultra uHPLC system equipped with a LEAP HTC PAL autosampler. Mobile phase A contains 0.01% acetic acid in 10 mM NH ₄ OAc and ACN / water (10:90) and mobile phase B is 0 in 10 mM NH ₄ OAc and ACN / water (90:10). Contains 0.1% acetic acid. Chromatographic separation is performed using an Acquity (R) uHPLC BEH C18 column (1.7 [mu] m, 2.1 x 50 mm) with 0-1.5 composed of 28% mobile phase B for the analysis of compound (I). Isocratic eluent in minutes, followed by a gradient eluent composed of a linear increase of 28% B to 100% B in 0.1 minutes, followed by holding it at 100% B for 1.1 minutes to flush the column Was done by. Subsequently, the gradient was returned to 28% B within 0.1 minutes and held at 28% for 0.9 minutes with a flow time of 3.7 minutes. The flow rate was 0.6 mL / min, and the column temperature was kept at 60 ° C. Detection was carried out using an AB Sciex Triple Quad 5500 mass spectrometer with turbo ion spray ionization with positive ESI and using multiple reaction monitoring (MRM) mode. The MRM transition was m / z 535 → 256 for compound (I) and m / z 541 → 256 for [ ¹³ C ₂ , D ₄ ] -compound (I). Data collection and quantification was performed using AB Sciex Analyst® 1.5.1 software. The calibration curve was 0.500 to 500 ng / ml for Compound (I) and fitted to a 1 / x2 weighted linear regression model. During sample analysis, the four levels of analytical quality control (QC) samples showing low, geometric mean, medium and high concentrations of Compound (I) prepared in human EDTA plasma are 4 at each concentration level for each analytical flow. Analyzed repeatedly. The results from these QC samples are used to accept or not accept flow analytes containing experimental samples based on preset acceptance criteria for diffraction of compound (I) in human EDTA plasma.

The results of this experiment are shown in Table 7 and FIG. A significant decrease in the AUC and Cmax of Compound (I) was seen in humans treated with 40 mg famotidine compared to pre-treated humans.
Table 7

  In vivo experiments were performed comparing the pharmacokinetics of the free base compound (I) and the hemisulfate of compound (I) in dogs treated with famotidine or pentagastrin.

Capsules containing 150 mg of the free base or compound (I) as hemisulfate were prepared:
1. Compound (I) free base capsule: 50% by weight of Compound (I), 42% by weight of microcrystalline cellulose, 3% by weight of croscarmellose sodium, 4% by weight of Klucel EXF hydroxypropylcellulose, 0.5% by weight Of magnesium stearate, 0.5% by weight of colloidal silicon dioxide. Compound (I) Hemisulfate Capsules: 57% Example Compound 1 (Compound (I) Hemisulfate, Crystalline Form H1.5-1), 40% Microcrystalline Cellulose, 3% Croscarmellose Sodium .

Four male dogs (10 kg) were treated according to the following three treatment protocols:
Treatment 1: Pretreatment with pentagastrin (6 μg / kg, IP) several hours prior to oral administration of Compound (I) free base capsule.
Treatment 2: Pre-orally with 40 mg famotidine 3 hours prior to oral administration of Compound (I) free base capsule.
Treatment 3: Pre-oral treatment with 40 mg famotidine 3 hours prior to oral administration of Compound (I) hemisulfate capsules.

Blood samples are collected at 0, 0.5, 1, 2, 4, 8, and 24 hours after administration of Compound (I) free base capsules or Compound (I) hemisulfate capsules and stored in EDTA tubes did. Compound (I) in canine EDTA plasma was analyzed using liquid-liquid extraction and uHPLC-MS / MS detection on a Triple Quad 5500 mass spectrometer. An aliquot of 0.050 mL of canine EDTA plasma was used for the assay. This method utilized [ ¹³ C ₂ , D ₄ ] -compound (I) labeled with a stable isotope as an internal standard. MeOH: 50 μL of 200 ng / mL [ ¹³ C ₂ , D ₄ ] -compound (I) in water (20/80) and 50 μL of 1M NH ₄ OAc containing 4% acetate buffer solution, 0.050 mL In addition to each experimental sample, quality control (QC) sample, and calibration standards, the sample was extracted by shaking with 600 μL of methyl tert-butyl ether (MTBE) for 15 minutes. 450 μL of the organic layer was removed and evaporated to dryness. The residue was reconstituted with 300 μL of reconstitution solution (30% acetonitrile in 10 mM NH ₄ OAc containing 0.01% acetic acid). All liquid transfer steps were performed using a Perkin Elmer JANUS Mini® liquid handler, except when adding an internal standard solution. An aliquot of 5 μL of the extracted sample was injected into the uHPLC-MS / MS system. This uHPLC was performed using a LEAP 4X Ultra uHPLC system equipped with a LEAP HTC PAL autosampler. Mobile phase A contains 0.01% acetic acid in 10 mM NH ₄ OAc and ACN / water (10:90) and mobile phase B is 0 in 10 mM NH ₄ OAc and ACN / water (90:10). Contains 0.1% acetic acid. Column chromatographic separation is performed using an Acquity® uHPLC BEH C18 column (1.7 μm, 2.1 × 50 mm) consisting of 28% mobile phase B for analysis of compound (I) 0-1. Hold the isocratic eluent at 5 minutes, followed by a gradient eluent composed of a linear increase from 28% B to 100% B in 0.1 minutes, and then hold at 100% B for 1.1 minutes to flush the column Was done by. The gradient was then returned to 28% B within 0.1 minutes and held at 28% for 0.9 minutes with a total flow time of 3.7 minutes. The flow rate was 0.6 mL / min, and the column temperature was kept at 60 ° C. Detection was performed with multiple reaction monitoring (MRM) mode using an AB Sciex Triple Quad 5500 mass spectrometer with turbo ion spray ionization with positive ESI. The MRM transition was m / z 535 → 256 for compound (I) and m / z 541 → 256 for [ ¹³ C ₂ , D ₄ ] -compound (I). Data collection and quantification was performed using AB Sciex Analyst® 1.5.1 software. The calibration curve ranged from 3.00 to 3000 ng / mL for compound (I) and was fitted to a 1 / x2 weighted linear regression model. During sample analysis, the four levels of analytical quality control (QC) samples showing low, geometric mean, medium and high concentrations of Compound (I) prepared in human EDTA plasma are 4 at each concentration level for each analytical flow. Analyzed repeatedly. The results from these QC samples are used to accept or not accept flow analytes containing experimental samples based on preset acceptance criteria for diffraction of compound (I) in human EDTA plasma.

The results of this experiment are shown in Table 8 and FIG. There was a significant decrease in AUC and Cmax in dogs treated with free base compound (I) and then pretreated with famotidine (high gastric pH) compared to dogs pretreated with pentagastrin (low gastric pH). . Administration of Example Compound 1 (hemisulfate sesquihydrate hydrate salt of Compound (I)) to dogs pre-treated with famotidine showed a too low decrease in AUC and Cmax. In this particular experiment, the hemisulfate salt of Compound (I), when pre-treated with famotidine, had a C _max value of 2596 ng / mL, 12473 ng · h / mL AUC _0-24hr , and 34.73. % Bioavailability. In contrast, in similar experiments, Compound (I) free base, when administered after pretreatment with famotidine, had a C _max value of 245 ng / mL, an AUC _{0-24 hr of 1762} ng · h / mL, and 4. Provided 54% bioavailability.
Table 8
Pharmacokinetic parameters for administration of 150 mg dogs of compound (I) as hemisulfate or free base

The hemisulfate salt of compound (I) has been found to be particularly advantageous compared to compound (I) free base. The hemisulfate salt of Compound (I) has the surprising benefit to patients of reducing fluctuations in the bioavailability of Compound (I) and / or increasing the bioavailability of Compound (I). Based on these in vitro and in vivo data, hemisulfate is expected to provide significant benefits for consistency in patient bioavailability over free form. As illustrated, in patient populations administered drugs that can increase the pH of gastric acid, such as antacids, proton pump inhibitors, or H ₂ -receptor antagonists, hemisulfate forms are significantly increased in biologics. Provide availability.

Single crystal data (LVL)
Data was collected on a Bruker-Nonius CAD4 continuous diffractometer. Unit cell parameters were obtained by least squares analysis of experimental diffractometer settings for 25 high-angle reflections. Intensities were measured using a CuKα ray (λ = 1.5418Å) at a constant temperature with a θ-2θ variable scan technique and corrected only for the Lorentz polarization factor. Background counts were collected during a scan that was half the scan time. Alternatively, single crystal data was collected on a Bruker-Nonius Kappa CCD2000 system using CuKα radiation (λ = 1.5418). The index and processing of the measured intensity data can be found in the HKL2000 software package (Otwinowski, Z & Minor, W. (1997) in Macromolecular Crystallography, eds. Carter, WC Jr. & Sweet, RM in the Collect program suite. (Academic, NY), Vol. 276, pp307-326). (Collect Data collection and processing user interface: Collect: Data collection software, R. Hooft, Nonius BV, 1998). Alternatively, single crystal data was collected on a Bruker-AXS APEX2 CCD system using CuKα radiation (λ = 1.5418). The measured intensity data was indexed and processed using the APEX2 software package / program suite (APEX2 Data collection and processing user interface: APEX2 User Manual, v1.27; BRUKER AXS, Inc., Madison, Wis.).

  Where indicated, crystals were cooled with a cryogenic stream of Oxford cryosystem (Oxford Cryosystems Cryostream cooler: J. Cosier and A.M. Glazer, J. Appl. Cryst., 1986, 19, 105) during data collection.

  SDP software package (SDP, Structure Determination Package, Enraf-Nonius, Bohemia NY) in which these structures are dissolved by a direct method and modified in part, or crystallographic package MAXUS (maXus solution and refinement software suite : S. Mackay, CJ Gilmore, C. Edwards, M. Tremayne, N. Stewart, K. Shankland. MaXus: a computer program for the solution and refinement of crystal structures from diffraction data or SHELXTL (Sheldrick, GM. 1997, SHELXTL. Structure Determination Programs. Version 5.10 or greater, Bruker AXS, Madison, Wisconsin).

The resulting atomic parameters (coordinates and temperature factors) were refined by a full matrix least squares method. The function minimized by refinement was Σ _w (| F _o | − | F _c |) ² . R _w = [Σ _w (| F _o | − | F _c |) ² / Σ _w | F _o | ² ] ^{1/2 where} w is an appropriate weighting function based on the error in the measured intensity. R] is defined as Σ || F _o | − | F _c || / Σ | F _o |. Difference maps were tested at all stages of refinement. Hydrogen was introduced into the theoretical position using an isotropic temperature factor, but the hydrogen parameters did not change.

Single crystal data (WFD)
Diffraction data was collected at room temperature using a Bruker SMART 2K CCD diffractometer equipped with a graphite monochromator emitting CuKα radiation (λ = 1.54056４０). The entire data set was collected using the ω scan mode over a 2θ range with a crystal-detector distance of 4.98 cm. For empirical absorption correction, a SADABS routine equipped with a diffractometer was used (Bruker AXS. 1998, SMART and SAINTPLUS. Area Detector Control and Integration Software, Bruker AXS, Madison, Wisconsin, USA). Final unit cell parameters were determined using the entire data set.

All structures were dissolved by direct methods and refined by full matrix least squares technique using the SHELXTL software package (Sheldrick, GM. 1997, SHELXTL. Structure Determination Programs. Version 5.10, Bruker AXS, Madison, Wisconsin, USA.). The function minimized by refinement was Σ _w (| F _o | − | F _c |) ² . R _w = [Σ _w (| F _o | − | F _c |) ² / Σ _w | F _o | ² ] ^{1/2 where} w is an appropriate weighting function based on the error in the measured intensity. R) is defined as Σ || F _o | − | F _c || / Σ | F _o |. Difference Fourier maps were tested at all stages of refinement. All non-hydrogen atoms were refined with anisotropic thermal displacement parameters. Hydrogen atoms bonded by hydrogen bonds were placed in the final difference Fourier map, and the positions of other hydrogen atoms were calculated from the theoretical structure using standard bond lengths and angles. They are assigned an isotropic temperature factor and are included in the calculation of structure factors using fixed parameters.

PXRD (Philips)
About 200 mg was packed into a Philips powder X-ray diffraction (PXRD) sample holder. The sample was transferred to a Philips MPD unit (45 KV, 40 mA, CuKα). Data is from 2 to 32 2-theta range (continuous scan mode, scan rate 0.03 degrees / second, auto divergence and anti scatter slit, receiving slit: 0.2 mm , Sample spinner (ON).

PXRD (GADDS-NB)
X-ray powder diffraction (PXRD) data was acquired using a Bruker C2 GADDS. The radiation was CuKα (40 KV, 40 mA). The sample-detector distance was 15 cm. The powder sample was placed in a sealed glass capillary with a diameter of 1 mm or less; the capillary was rotated during data collection. Data was collected for 3 ≦ 2θ ≦ 35 ° with a sample exposure time of at least 1000 seconds. The resulting two-dimensional diffraction arcs were integrated to create a conventional one-dimensional PXRD pattern with a step size of 0.02 degrees 2θ in the range of 2 to 35 degrees 2θ.

DSC (Open Pan)
Differential scanning calorimetry (DSC) experiments were performed on TA instruments® model Q2000, Q1000 or 2920. Samples (approximately 2-6 mg) were weighted with an aluminum pan, accurately recorded to 1/100 milligram and transferred to the DSC. The instrument was purged with nitrogen gas at 50 mL / min, data was collected between room temperature and 300 ° C. at a heating rate of 10 ° C./min. The plot was made with a reduced endothermic peak.

TGA (open bread)
Thermogravimetric analysis (TGA) experiments were performed on TA Instruments model Q500 or 2950. A sample (about 10-30 mg) was placed on a pre-weighed platinum pan. The weight of the sample was accurately measured and recorded to 1/1000 milligram with the instrument. The furnace was purged with nitrogen gas at 100 mL / min. Data was collected between room temperature and 300 ° C. at a heating rate of 10 ° C./min.

Solid-state nuclear magnetic resonance (SSNMR)
All solid C-13 NMR measurements were performed using a Bruker AV-400, 400 MHz NMR spectrometer. High resolution spectra include high power proton decoupling and ramp amplitude cross-polarization (RAMP-CP) with TPPM pulse sequence and magic-angle spinning (MAS) of about 12 kHz. (AE Bennett et al, J. Chem. Phys., 1995, 103, 6951), (G. Metz, X. Wu and SO Smith, J. Magn. Reson. A ,. 1994, 110, 219-227). Approximately 70 mg of sample was packed into a canister-design zirconia rotor and used for each experiment. The chemical shift (δ) was based on an external reference adamantane with high frequency resonance set at 38.56 ppm (WL Earl and DL VanderHart, J. Magn. Reson., 1982, 48, 35-54).

VTI (dry on)
Hygroscopic isotherms were collected on a VTI SGA-100 Symmetric Vapor Analyzer using approximately 10 mg of sample. The sample was dried at 60 ° C. for 10 minutes until a loss rate of 0.0005 wt% / min was obtained. The samples were tested at 25 ° C. and 3 or 4, 5, 15, 25, 35, 45, 50, 65, 75, 85, and 95% RH. Equilibrium at each RH was achieved at a rate of 0.0003 wt% / min for 35 minutes or up to 600 minutes.

  It will be apparent to those skilled in the art that the present disclosure is not limited to the exemplary embodiments described above and may be embodied in other specific forms without departing from the required aspects thereof. Therefore, the examples should be considered as illustrative and not as limiting, and refer to the claims, not the examples, and thus the meaning of the claims. All changes within the range should be acceptable to the present invention.

Claims (13)

Compound (I):

(I)
A pharmaceutical composition comprising 70 mg to 80 mg of a hemisulfate salt and a pharmaceutically acceptable carrier or diluent. The pharmaceutical composition according to claim 1 , comprising 70 mg of hemisulfate of compound (I). The pharmaceutical composition according to claim 1 , comprising 80 mg of the hemisulfate of compound (I). The pharmaceutical composition according to any one of claims 1 to 3 , wherein the hemisulfate of compound (I) is sesquihydrate. The hemisulfate of compound (I) is:
following:
Lattice size: a = 10.92Å
b = 33.04Å
c = 7.90cm
α = 90 degrees
β = 90 degrees
γ = 90 degree space group: P2 ₁ 2 ₁ 2
Compound (I) molecule / asymmetric unit: 1
Volume = 2851 ^{３ 3}
Density (theoretical value) = 1.423 g / cm ³
(Measurement of the crystal form is at a temperature of 25 ° C.);
The pharmaceutical composition according to any one of claims 1 to 4 , which is in crystalline form H1.5-1 characterized by unit cell parameters equivalent to. The form H1.5-1 is:
a) Figure 1:

Measured powder X-ray diffraction pattern according to the pattern shown in FIG.
b) Figure 1:

A simulated powder X-ray diffraction pattern with the pattern shown in FIG.
c) 5.4 ± 0.1, 8.6 ± 0.1, 9.7 ± 0.1, 12.4 ± 0.1, 14.9 ± 0.1, 17.6 ± 0.1, 18. Powder X-ray diffraction comprising 4 or more 2θ values selected from 18.1 ± 0.1, 20.5 ± 0.1, 21.4 ± 0.1, and 22.0 ± 0.1 Pattern (CuKα λ = 1.5418Å) (measurement of crystal form is at a temperature of 25 ° C.); .1, 30.7 ± 0.1, 43.1 ± 0.1, 45.9 ± 0.1, 47.1 ± 0.1, 52.0 ± 0.1, 54.2 ± 0.1 72.5 ± 0.1, 117.0 ± 0.1, 117.7 ± 0.1, 124.2 ± 0.1, 125.2 ± 0.1, 128.3 ± 0.1, 130 .3 ± 0.1, 131.4 ± 0.1, 134.1 ± 0.1, 140. 8 ± 0.1, 144.7 ± 0.1, 148.7 ± 0.1, 149.8 ± 0.1, 151.2 ± 0.1, 153.4 ± 0.1, 155.1 ± Solid nuclear magnetic spectrum containing 6 or more peaks (δ (ppm) normalized by TMS) selected from 0.1, 155.6 ± 0.1, and 156.7 ± 0.1:
Characterized by one or more of:
The pharmaceutical composition according to claim 5 . The pharmaceutical composition according to any one of claims 1 to 6 , for use in inhibiting CGRP receptors. The pharmaceutical composition according to any one of claims 1 to 6 , for use in the treatment of migraine. 7. A pharmaceutical composition according to any one of claims 1 to 6 for oral administration useful for the treatment of migraine. The pharmaceutical composition according to any one of claims 1 to 6 , for sublingual administration useful for the treatment of migraine. 7. A pharmaceutical composition according to any one of claims 1 to 6 for intranasal administration useful for the treatment of migraine. The pharmaceutical composition according to any one of claims 1 to 6 , for use once a day in the treatment of migraine. The pharmaceutical composition according to any one of claims 1 to 6 , for use twice a day in the treatment of migraine.

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